ORIGINAL RESEARCH
published: 14 March 2017
Edited by:
Axel Cloeckaert,
Institut National de la Recherche
Agronomique, France
Reviewed by:
Samus Fanning,
University College Dublin, Ireland
Iddya Karunasagar,
Nitte University, India
*Correspondence:
Bruno R. Pribul
bpribul@gmail.com
Specialty section:
This article was submitted to
Antimicrobials, Resistance and
Chemotherapy,
a section of the journal
Frontiers in Microbiology
Received: 29 September 2016
Accepted: 14 February 2017
Published: 14 March 2017
Citation:
Pribul BR, Festivo ML, Rodrigues MS,
Costa RG, Rodrigues ECdP,
de Souza MMS and Rodrigues DdP
(2017) Characteristics of Quinolone
Resistance in Salmonella spp. Isolates
from the Food Chain in Brazil.
Front. Microbiol. 8:299.
doi: 10.3389/fmicb.2017.00299
Frontiers in Microbiology | www.frontiersin.org
doi: 10.3389/fmicb.2017.00299
Characteristics of Quinolone
Resistance in Salmonella spp.
Isolates from the Food Chain in Brazil
Bruno R. Pribul 1, 2*, Marcia L. Festivo 1 , Marcelle S. Rodrigues 1 , Renata G. Costa 1 ,
Elizabeth C. dos P. Rodrigues 1 , Miliane M. S. de Souza 2 and Dalia dos P. Rodrigues 1
1
National Reference Laboratory for Enteric Diseases, Oswaldo Cruz Institute(FIOCRUZ), Rio de Janeiro, Brazil, 2 Laboratory
of Veterinary Bacteriology, Federal Rural University of Rio de Janeiro, UFRRJ, Rio de Janeiro, Brazil
Salmonella spp. is an important zoonotic pathogen related to foodborne diseases.
Despite that quinolones/fluoroquinolones are considered a relevant therapeutic strategy
against resistant isolates, the increase in antimicrobial resistance is an additional difficulty
in controlling bacterial infections caused by Salmonella spp. Thus, the acquisition of
resistance to quinolones in Salmonella spp. is worrisome to the scientific community
along with the possibility of transmission of resistance through plasmids. This study
investigated the prevalence of plasmid-mediated quinolone resistance (PMQR) in
Salmonella spp. and its association with fluoroquinolone susceptibility in Brazil. We
evaluated 129 isolates, 39 originated from food of animal sources, and 14 from
environmental samples and including 9 from animals and 67 from humans, which were
referred to the National Reference Laboratory of Enteric Diseases (NRLEB/IOC/RJ)
between 2009 and 2013. These samples showed a profile of resistance for the tested
quinolones/fluoroquinolones. A total of 33 serotypes were identified; S. Typhimurium
(63) was the most prevalent followed by S. Enteritidis (25). The disk diffusion test
showed 48.8% resistance to enrofloxacin, 42.6% to ciprofloxacin, 39.53% to ofloxacin,
and 30.2% to levofloxacin. According to the broth microdilution test, the resistance
percentages were: 96.1% to nalidixic acid, 64.3% to enrofloxacin, 56.6% to ciprofloxacin,
34.1% to ofloxacin, and 30.2% to levofloxacin. Qnr genes were found in 15 isolates
(8 qnrS, 6 qnrB, and 1 qnrD), and the aac(6 )-Ib gene in 23. The integron gene was
detected in 67 isolates with the variable region between 600 and 1000 bp. The
increased detection of PMQR in Salmonella spp. is a serious problem in Public Health
and must constantly be monitored. Pulsed-field gel electrophoresis was performed to
evaluated clonal profile among the most prevalent serovars resistant to different classes
of quinolones. A total of 33 pulsotypes of S. Typhimurium were identified with a low
percentage of genetic similarity (65%). This result demonstrates the presence of high
diversity in the resistant clones evaluated in this study.
Keywords: foodborne diseases, Salmonella spp., quinolone resistance, plasmid mediated quinolone resistance,
clonal profile
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Pribul et al.
INTRODUCTION
Foodborne diseases caused by Salmonella spp. are a serious
public health problem in many parts of the world. The variety
of food sources, particularly foods of animal origin, and routes of
transmission can lead to human infection (Scallan et al., 2011).
In addition, the progressive increase of antimicrobial
resistance in foodborne Salmonella isolates is observed as due
to the uncontrolled use of these drugs for therapeutic and
prophylactic purposes in foods of animal origin such as poultry,
pigs, and cattle. These events have reinforced the need for
epidemiological studies describing the prevalence and patterns
of resistance in these bacteria (Yang et al., 2010; Tamang et al.,
2011). Antimicrobial-resistant bacteria emerge from the use of
antimicrobial drugs to treat and prevent diseases and promote
growth in large-scale animal production.
Quinolones, particularly fluoroquinolones, are commonly
used for the treatment of multi-drug resistant salmonellosis in
human and veterinary medicine because of their broad spectrum
antimicrobial activity (Dalhoff, 2012).
Point mutations in DNA gyrase and topoisomerase IV genes
are directly related to quinolone resistance in Enterobacteriaceae
by changes in the action target site called quinolone resistance-
determining regions (QRDR). In Salmonella spp., these
mutations are related to resistance to nalidixic acid (NAL) and
reduced susceptibility to FQs such as that of ciprofloxacin (Cip)
(Cavaco and Aarestrup, 2009). It is believed that the resistance
to quinolones is mediated only by this mechanism. However, the
situation changed with the discovery of a variety of determinants
of plasmid-mediated quinolone resistance (PMQR).
Currently three mechanisms are recognized as PMQRs. The
qnr genes with five different qnr families, each with different
numbers of alleles (qnrA17, qnrS14, qnrB131, qnrC, and
qnrD) (Jacoby et al., 2009); a modified aminoglycoside acetyl-
transferase gene [aac(6 )-1b-cr] (Robicsek et al., 2005); and a
specific quinolone efflux pump (qepA) (Yamane et al., 2007)
R
and multidrug resistance pumps such as oqxAB (Zhao et al.,
2010). PMQR-positive isolates present a low-level of resistance to
quinolones (only a small reduction in susceptibility to nalidixic
acid). However, the ability to highlight pre-existing resistance
mechanisms, such as chromosomal mutations in target regions
of quinolones that still allow the selection of resistant mutants
to quinolone concentrations (therapeutic doses), emphasize the
importance of studying these genes (Cui et al., 2014).
The present study identified the occurrence of some PMQR in
Salmonella spp. isolated between 2009 and 2013 from the food
chain in Brazil, and characterized the genetic similarity profile of
serovars of greatest importance in the dispersion of resistance to
quinolones in Brazil.
MATERIALS AND METHODS
Bacterial Isolates
129 Salmonella spp. strains with resistance to quinolone and/or
fluoroquinolone were evaluated. Of these, 51.9% (67/129) were
from human clinical isolates, 30.2% (39/129) from food products
for human consumption (beef, eggs, milk), 7.1% (9/129) from
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Quinolone Resistance in Salmonella spp. in Brazil
food of animal origin for human consumption (poultry, swine,
cattle), and 10.8% (14/129) from environmental samples (water
and drag swabs); all samples were selected from a database.
The studied strains were sent to the National Reference
Laboratory of Enteric Diseases (NRLEB/IOC/RJ) between 2009
and 2013 and stored in phosphate-buffered agar at room
temperature and/or in BHI/glycerol broth 70 C. The isolates
were inoculated in Nutrient Broth (DIFCO) and incubated
at 37 C for 1218 h for subsequent tests, as confirmation
of the biochemical, serological and antimicrobial resistance
profile.
Antigenic Characterization
The serological determination of Salmonella serotypes was
determined according to the Kauffmann-White scheme using
slide agglutination with O and H antisera prepared in the
LRNEB/IOC/RJ.
Antimicrobial Susceptibility
The obtained resistance profiles were confirmed by the disk
diffusion test according to Clinical and Laboratory Standards
Institute (2013, 2014). According to Pribul et al. (2016), this
test was performed using representatives of the quinolone class
(OXOID) for human and veterinary therapeutic use, such as
Nalidixic Acid (NAL), Ciprofloxacin (CIP), Enrofloxacin (ENO),
Ofloxacin (OFL), and Levofloxacin (LVX).
MIC determinations for Nalidixic Acid (SIGMA),
Ciprofloxacin (SIGMA), Enrofloxacin (SIGMA), Levofloxacin
(SIGMA), and Ofloxacin (SIGMA) were performed in
96-well-microplates and according to the Clinical and Laboratory
Standards Institute (2013) broth microdilution assay.
Detection of PMQR
Total DNA was extracted using the DNEASY Tissue Qiagen
kit. The studied genes were detected by PCR amplification using
primer sequences reported in Pribul et al. (2016). The qnrA,
qnrB, and qnrS genes were amplified through multiplex PCR
reactions; the rrs gene was used as the reaction control. The qnrC,
qnrD, aac(6 )-Ib, integrase, and variable integron region genes
were amplified by simplex PCR.
PFGE
The isolates from serovars S. Typhimurium, S. Muenchen,
S. Infantis, and S. Heidelberg were subjected to molecular typing
by pulsed-field gel electrophoresis, which clonally evaluates
isolates. The PulseNet protocol was used in this study including
DNA preparation according to Heir et al. (2000) and digestion
with XbaI restriction enzyme, according to Pfaller et al.
(1992), Tenover et al. (1997), and Cooper et al. (2006). The
definition of clones was based on the recommendations of
Tenover et al. (1997) and Barrett et al. (2006). S. Braenderup
H9812, which is considered the universal strain for PulseNet
(Hunter et al., 2005), was used as the standard. The restriction
patterns were analyzed in the BioNumerics software IV (Applied
Maths).
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Pribul et al.
RESULTS
Serovar Identified
Altogether, 26 different Salmonella serovars were identified.
Salmonella Typhimurium (48.8%, 63/129) was the predominant
serovar followed by Salmonella Enteritidis (19.4%, 25/129). The
prevalent serovars associated with resistance to quinolones are
presented in Table 1.
Most of the studied samples were isolated in 2012 (88 of 129).
Among these 129 isolates that were previously resistant
to Nalidixic Acid, five were sensitive to all tested quinolones
(including Nalidixic Acid), 55 (42.6%) were resistant to
Ciprofloxacin, 63 (48.8%) to Enrofloxacin, 51 (39.53%) to
Ofloxacin, and 48 (37.2%) to Levofloxacin in the disc diffusion
test.
The broth microdilution test identified 36.4% (47/129)
isolates with decreased susceptibility to Ciprofloxacin (MICs
between 0.125 and 0.5 mg/ml), 20.1% (26/129) to Enrofloxacin,
9.3% (12/129) to Ofloxacin, and 6.2% (8/129) to Levofloxacin
(MICs between 0.5 and 1 mg/ml). The decreased susceptibility
breakpoint to Nalidixic Acid is not reported by Clinical
and Laboratory Standards Institute (2015). Seventy-three
(56.6%) isolates were resistant to Ciprofloxacin, 83 (64.3%)
to Enrofloxacin, 44 (34.1%) to Ofloxacin, and 39 (30.2%) to
Levofloxacin. A total of 124 isolates (96.1%) were resistant to
Nalidixic Acid.
The resistance profile obtained with the microdilution test
showed that 37 (28.7%) isolates were resistant to all tested
quinolones, 30 (23.2%) to Ciprofloxacin, Enrofloxacin, and
Nalidixic Acid, 16 (12.4%) to Enrofloxacin and Nalidixic Acid,
2 (1.5%) to Ciprofloxacin and Nalidixic Acid, and 39 (30.2%) to
Nalidixic Acid only.
The detection of resistance genes showed six isolates carrying
the qnrB gene, eight the qnrS gene, and one the qnrD gene.
Among these 15 positive isolates, 10 strains were recovered
from human samples, 3 from food of animal origin, 1 from
environmental samples, and 1 from animal samples. The most
qnr-positive prevalent serovar was S. Typhimurium followed by
S. Saintpaul and S. Livingstone. None of the isolates presented the
qnrA or qnrC genes.
A total of 23 isolates showed the aac(6 )-Ib gene, which is
prevalent in S. Typhimurium (14/23). The most prevalent source
of isolation was human (10/23), followed by foodborne (7/23),
animal (3/23), and amibental (3/23). Thirteen isolates aac(6 )-Ib
positive were resistant to all tested quinolones.
Three qnr-positive isolates presented the aac(6 )-Ib gene in
association: two S. Typhimurium and one S. Saintpaul. These
two Salmonella ser. Typhimurium were resistant to all tested
quinolones/fluoroquinolones in the broth microdilution assay at
the highest concentration.
Sixty-seven isolates showed the presence of integrase gene
within 600 to 1000 bp variable region range and were
mainly identified in human samples (38/67) followed by food
samples (15/67), ambiental (8/67), and animal (6/67). The
S. Typhimurium serovar was the most frequent (39/67) isolate
with the conserved region of class 1 integron and variable
regions between 900 and >1000 bp. Nine isolates of serovar
S. Typhimurium carrying the aac(6 )-Ib gene were positive for
Frontiers in Microbiology | www.frontiersin.org
Quinolone Resistance in Salmonella spp. in Brazil
TABLE 1 | Distribution of quinolone-resistant Salmonella spp. serovars
isolated from food chain diseases.
Salmonella spp. Serotype Number of NTSa from
Human Food Environment animal Total
S. Typhimurium 35 22 4 2 63
S. Enteritidis 24 1 25
S. Muenchen 2 2 4
S. Infantis 1 1 3
S. Heidelberg 2 1 3
Others 3 11 8 5 26
Total 67 37 12 8
a Non-typhoidal Salmonella.
the integron with 900 bp; two of these were also positive for
qnr.
Figure 1 shows the clonal profile comparison between the
quinolones/fluoroquinolones resistant strains and other sensitive
strains.
The genetic similarity among isolates with resistance to
quinolones was approximated 84% in S. Infantis, 92% in
S. Heidelberg, 88% in S. Muenchen, and 63% in S. Typhimurium
despite their isolation in different periods, regions, and sources.
Thirty-three distinct pulsotypes were identified among strains
with low percentage genetic similarity in serovar S. Typhimurium
(65%), representing the highest diversity among resistant
clones.
The Table 2 presents resistance profiles obtained with the
microdilution test, detection of PMQR, size of variable integron
region, and the pulsotype identified by the pfge technique.
DISCUSSION
The variation in resistance to the different tested quinolones can
be explained by the mechanism of resistance when the resistance
level depends on the affected target enzyme, the number of
accumulated mutations, and presence of PMQRs. Furthermore,
there is a relationship between the level of specific resistance
and potency of each drug, especially in the newest quinolones
(Sanders, 2001; Ruiz et al., 2012).
Chong et al. (2010) reported that an increased resistance to
fluoroquinolones based on the acquisition of qnr genes could
be related with reduction in the clinical efficacy of this class
of antimicrobial. However, Jacoby et al. (2009) argue that the
genes involved in plasmid-level resistance to fluoroquinolones
are still poorly understood when compared to other resistance
mechanisms.
A high prevalence of isolates carrying PRQM genes is
reported in the present study (27%, 35/129). The most prevalent
serovar associated with the presence of PMQR genes was
Salmonella ser. Typhimurium (18/35). A high level of detection
of S. Typhimurium was expected because this serovar is
directly related to detected genotypic and phenotypic profiles
of antimicrobial resistance (Herrero et al., 2008; Kingsley et al.,
2009). The presence of PMQR genes is related to decreased
susceptibility to fluoroquinolones, accelerating the selection of
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Pribul et al.
FIGURE 1 | Distribution of phylogenetic groups of Salmonella ser.
Infantis, Heidelberg, Muenchen, and Typhimurium according to PFGE. Isolates
with resistance to quinolones; a ID of IOC/year of isolation; b region; c state;
d source of isolation; e serovar; f pulsotype.
Frontiers in Microbiology | www.frontiersin.org
Quinolone Resistance in Salmonella spp. in Brazil
fluoroquinolone-resistant mutants (Rodrguez-Martnez et al.,
2011).
Three isolates presented association between the qnr and
aac(6 )-Ib genes. A similar association has been reported by
Park et al. (2006) in the United States, Xiong et al. (2011) while
investigating the aac(6 )-Ib and qnr genes in Enterobacter cloacae
in China, and Kim et al. (2013) in enterobacteria isolated from
clinical samples in Korea. Not sequencing the aac(6 )-Ib gene to
determine the cr variant was one limitation in the present study.
Regardless that some authors recognize the location of the
aac(6 )-Ib gene mostly in class 1 integrons, our results show the
absence of this gene in all analyzed strains [12 aac(6 )-Ib positive
isolates without the integron region] (Rodrguez-Martnez et al.,
2011; Kim et al., 2013).
The Enteritidis serovar was not assessed by PFGE because,
according to the literature, these isolates have low clonal diversity
(Spiliopoulou et al., 2007).
Six distinct pulsotypes were detected in S. Infantis serovar
isolates. Those with resistance to quinolones are placed in two
separate pulsotypes (BRJFXX01.13 and BRJFXX01.12) with a
genetic similarity of 85%. The quinolone resistance isolates
were obtained from different sources, regions, and periods, and
the resistance to quinolones showed variations. The 6754/12
isolate showed resistance to ciprofloxacin, nalidixic acid, and
enrofloxacin and carried the qnrD gene; the 9606/10 isolate
showed resistance to nalidixic acid only and did not carry
resistant genes.
The S. Heidelberg serovar presented eight distinct pulsotypes.
The resistant isolates showed a clonal ratio of 100% similarity
between the isolates 5/12 and 19/12, and 94% between them
and isolate 11394/11. The 11394/11 isolate (environmental
source from the Southern region) was detected in the
BRJF6X01.004 pulsotype (pulsotype with 13 susceptible isolates).
Isolates 5/12 and 19/12, within the same pulsotype, were
foodborne and originated in the Southern region.
Three pulsotypes were identified in serovar Muenchen isolates
with quinolone resistance profiles (pulsotypes BRJJ6X01008,
BRJJ6X01007, and BRJJ6X01006), showing 88% of genetic
similarity. Among the isolates resistant, the isolates of human
origin presenting a profile similarity of 97%. The isolates
of foodborne origin presenting the same origin clonal being
from different periods and states. The 2128/12, 2120/12, and
1192/12 isolates show similar quinolone resistance profiles.
However, isolate 851/11 show a resistance profile to ciprofloxacin
and ofloxacin. The 2120/12 and 2128/12 isolates show similar
resistance profile and are carriers of the qnrS gene.
The detection of 33 different pulsotypes of S. Typhimurium
indicates that different clones with resistance to quinolones
are circulating in Brazil. The most prevalent pulsotype
(BRJPXX01.090) is mainly represented in samples from the
Southern region and are related to food and human sources.
Most isolates of this pulsotype show the same resistance profile to
quinolones/fluoroquinolones (except isolates 3309/11, 5179/10,
591/12, and 667/12) demonstrating a resistance profile to all
tested quinolones/fluoroquinolones. Out of the 25 isolates
showing resistance to quinolones, 4 did not carry resistance
genes. Twelve isolates show an integron with a variable region of
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TABLE 2 | Resistance profile and resistance genes in isolates evaluated by PFGE.

Serovar Source IOC Pulsotype PMQRc Integron (bp) Resistance profile (MICb )
ID/Yeard

Infantis F 6754/12 BRJFXX01.13 qnrD CIP NAL ENOa

Infantis H 9606/10 BRJFXX01.012 NAL
Heidelberg F 19/12 BRJF0X01.024 700 CIP NAL ENO
Heidelberg F 5/12 BRJF0X01.024 CIP NAL ENO
Heidelberg H 11394/11 BRJF0X01.004 900 CIP NAL ENO
Muenchen H 2120/12 BRJJ6X01.008 qnrS NAL ENO
Muenchen H 2128/12 BRJJ6X01.007 qnrS NAL ENO
Muenchen F 1192/12 BRJJ6X01.006 aac(6 )-Ib 600 NAL ENO
Muenchen F 851/11 BRJJ6X01.006 - 700 CIP NAL ENO OFL
Typhimurium F 1618/12 BRJPXX01.095 aac(6 )-Ib CIP NAL ENO LVX OFL
Typhimurium H 5971/12 BRJPXX01.091 aac(6 )-Ib 1000 NAL
Typhimurium F 1310/12 BRJPXX01.093 qnrS NAL ENO
Typhimurium H 6744/12 BRJPXX01.089 qnrB 1000 NAL ENO
Typhimurium F 711/12 BRJPXX01.094 qnrD NAL
Typhimurium E 505/09 BRJPXX01.092 900 CIP NAL ENO
Typhimurium F 2263/12 BRJPXX01.096 CIP NAL ENO LVX OFL
Typhimurium A 2629/12 BRJPXX01.097 aac(6 )-Ib 900 CIP NAL ENO LVX OFL
Typhimurium F 14488/10 BRJPXX01.098 900 CIP NAL ENO
Typhimurium H 3307/12 BRJPXX01.080 1000 CIP NAL ENO
Typhimurium H 833/12 BRJPXX01.080 CIP NAL ENO
Typhimurium H 434/12 BRJPXX01.081 CIP NAL ENO
Typhimurium F 456/12 BRJPXX01.078 1000 NAL
Typhimurium H 5974/12 BRJPXX01.078 1000 NAL OFL
Typhimurium H 5977/12 BRJPXX01.078 900 CIP NAL ENO OFL
Typhimurium F 1622/12 BRJPXX01.077 NAL ENO
Typhimurium F 43/12 BRJPXX01.076 NAL
Typhimurium E 5973/12 BRJPXX01.076 1000 CIP NAL ENO
Typhimurium H 792/09 BRJPXX01.103 900 CIP NAL
Typhimurium E 12720/11 BRJPXX01.104 900 CIP NAL ENO LVX OFL
Typhimurium F 5417/10 BRJPXX01.105 900 CIP NAL ENO LVX OFL
Typhimurium H 17343/09 BRJPXX01.101 1000 CIP NAL ENO
Typhimurium H 2490/11 BRJPXX01.102 900 CIP NAL ENO LVX OFL
Typhimurium H 6627/11 BRJPXX01.089 900 CIP NAL ENO LVX OFL
Typhimurium H 19754/11 BRJPXX01.099 CIP NAL ENO LVX OFL
Typhimurium F 6161/11 BRJPXX01.099 CIP NAL ENO LVX OFL
Typhimurium H 744/12 BRJPXX01.099 900 CIP NAL ENO LVX OFL
Typhimurium H 1099/12 BRJPXX01.090 1000 CIP NAL ENO LVX OFL
Typhimurium H 12362/11 BRJPXX01.090 900 CIP NAL ENO LVX OFL
Typhimurium E 12722/11 BRJPXX01.090 CIP NAL ENO LVX OFL
Typhimurium F 129/12 BRJPXX01.090 aac(6 )-Ib CIP NAL ENO LVX OFL
Typhimurium H 13873/11 BRJPXX01.090 900 CIP NAL ENO LVX OFL
Typhimurium H 13874/11 BRJPXX01.090 900 CIP NAL ENO LVX OFL
Typhimurium F 14119/11 BRJPXX01.090 900 CIP NAL ENO LVX OFL
Typhimurium F 1615/12 BRJPXX01.090 CIP NAL ENO LVX OFL
Typhimurium F 16238/09 BRJPXX01.090 900 CIP NAL ENO LVX OFL
Typhimurium H 17242/11 BRJPXX01.090 900 CIP NAL ENO LVX OFL
Typhimurium H 2130/12 BRJPXX01.090 aac(6 )-Ib CIP NAL ENO LVX OFL
Typhimurium F 2178/12 BRJPXX01.090 aac(6 )-Ib CIP NAL ENO LVX OFL
Typhimurium F 2179/12 BRJPXX01.090 aac(6 )-Ib CIP NAL ENO LVX OFL

(Continued)

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Pribul et al. Quinolone Resistance in Salmonella spp. in Brazil

TABLE 2 | Continued

Serovar Source IOC Pulsotype PMQRc Integron (bp) Resistance profile (MICb )
ID/Yeard

Typhimurium F 2316/12 BRJPXX01.090 900 CIP NAL ENO LVX OFL

Typhimurium E 320/12 BRJPXX01.090 aac(6 )-Ib 900 CIP NAL ENO LVX OFL
Typhimurium H 3309/12 BRJPXX01.090 qnrB/aac(6 )-Ib 900 CIP NAL ENO
Typhimurium F 5179/10 BRJPXX01.090 CIP NAL ENO
Typhimurium F 591/12 BRJPXX01.090 900 NAL
Typhimurium H 5972/12 BRJPXX01.090 aac(6 )-Ib 900 CIP NAL ENO LVX OFL
Typhimurium F 667/12 BRJPXX01.090 CIP NAL ENO
Typhimurium H 6822/12 BRJPXX01.090 900 CIP NAL ENO LVX OFL
Typhimurium H 778/12 BRJPXX01.090 qnrB CIP NAL ENO LVX OFL
Typhimurium F 8796/10 BRJPXX01.090 900 CIP NAL ENO LVX OFL
Typhimurium A 8891/10 BRJPXX01.090 900 CIP NAL ENO LVX OFL
Typhimurium H 994/12 BRJPXX01.090 900 CIP NAL ENO OFL
Typhimurium H 5970/12 BRJPXX01.100 aac(6 )-Ib CIP NAL ENO LVX OFL
Typhimurium H 63/13 BRJPXX01.027 1000 CIP NAL ENO
Typhimurium H 6826/12 BRJPXX01.082 1000 NAL
Typhimurium H 55/13 BRJPXX01.073 qnrD CIP NAL ENO LVX OFL
Typhimurium H 5968/12 BRJPXX01.083 1000 NAL
Typhimurium H 6827/12 BRJPXX01.084 1000 NAL
Typhimurium H 431/12 BRJPXX01.086
Typhimurium H 5906/12 BRJPXX01.042 NAL
Typhimurium F 1199/09 BRJPXX01.075 600 CIP NAL ENO
Typhimurium H 5976/12 BRJPXX01.087 aac(6 )-Ib 700 CIP NAL ENO LVX OFL
Typhimurium H 777/12 BRJPXX01.088 qnrB/aac(6 )-Ib CIP NAL ENO LVX OFL

a CIP, Ciprofloxacin; ENO, Enrofloxacin; NAL, Nalidixic cid; LVX, Levofloxacina; OFL, Ofloxacin; b MIC, Minimum Inhibitory Concentration; c PMQR, Plasmid-Mediated Quinolone

Resistance; d IOC ID/Year, Institut Oswaldo Cruz Identification by Year; e F, Food; A, Animal; E, Environment; H, Human.

900 bp and one with >1000 bp. One isolate shows the qnrB gene
and, four show the aac(6 )-Ib gene. Two isolates show the 900 bp
integron and the aac(6 )-Ib gene; one isolate shows the 900 bp
integron and the qnrB, acc(6 )-Ib gene.
The profiles identified in the PFGE analysis show relatively
high diversity in all serovars and, indicate that cases of resistance
to quinolones are probably sporadic. This interpretation is in
accordance with other results reported in the literature (Feasey
et al., 2012).
AUTHOR CONTRIBUTIONS
All authors listed, have made substantial, direct and intellectual
contribution to the work, and approved it for publication.

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Conflict of Interest Statement: The authors declare that the research was
conducted in the absence of any commercial or financial relationships that could
be construed as a potential conflict of interest.
Copyright 2017 Pribul, Festivo, Rodrigues, Costa, Rodrigues, de Souza and
Rodrigues. This is an open-access article distributed under the terms of the Creative
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